Molecular characterization of potato fumarate hydratase and functional expression in Escherichia coli

Plant Physiology
G Nast, B Müller-Röber

Abstract

The tricarboxylic acid cycle enzyme fumarase (fumarate hydratase; EC 4.2.1.2) catalyzes the reversible hydration of fumarate to L-malate. We report the molecular cloning of a cDNA (StFum-1) that encodes fumarase from potato (Solanum tuberosum L.). RNA blot analysis demonstrated that StFum-1 is most strongly expressed in flowers, immature leaves, and tubers. The deduced protein contains a typical mitochondrial targeting peptide and has a calculated molecular mass of 50.1 kD (processed form). Potato fumarase complemented a fumarase-deficient Escherichia coli mutation for growth on minimal medium that contains acetate or fumarate as the sole carbon source, indicating that functional plant protein was produced in the bacterium. Antiserum raised against the recombinant plant enzyme recognized a 50-kD protein in wild-type but not in StFum-1 antisense plants, indicating specificity of the immunoreaction. A protein of identical size was also detected in isolated potato tuber mitochondria. Although elevated activity of fumarase was previously reported for guard cells (as compared with mesophyll cells), additional screening and genomic hybridization data reported here do not support the hypothesis that a second fumarase gene is expressed...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Jan 1, 1975·The Journal of General Virology·G VervlietJ Schell
Oct 5, 1990·Journal of Molecular Biology·S F AltschulD J Lipman
Jan 1, 1987·Annual Review of Biochemistry·P A Srere
Apr 1, 1989·European Journal of Biochemistry·G von HeijneR G Herrmann
Apr 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·W R Pearson, D J Lipman
May 31, 1988·Biochemical and Biophysical Research Communications·J C SacchettiniG A Grant

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Citations

Jul 27, 2001·Molecular Biology and Evolution·D GerbodE Viscogliosi
Mar 29, 2011·Phytochemistry·Wagner L AraújoAlisdair R Fernie
Apr 28, 2007·The Plant Journal : for Cell and Molecular Biology·Adriano Nunes-NesiAlisdair R Fernie
May 25, 2001·Journal of Experimental Botany·W H Outlaw, S Zhang
Jul 22, 2006·The Journal of Biological Chemistry·Virginie CoustouFrédéric Bringaud
Feb 8, 2021·International Journal of Biological Macromolecules·Iara Aimê CardosoMaria Cristina Nonato

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